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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 53 issue 7 (july 2019) : 874-879,   Doi: 10.18805/ijar.B-882

Expression and characterization of a lysine rich protein in cow milk

X
Xin Ma
L
Li Su
P
Peng Zhang
S
Sheng Zhang
B
Bo Tang
X
Xueming Zhang
W
Weimin Luan
Z
Ziyi Li
1College of Animal Science and Technology, Jilin Agricultural University, Changchun, China.
Cite article:- Ma Xin, Su Li, Zhang Peng, Zhang Sheng, Tang Bo, Zhang Xueming, Luan Weimin, Li Ziyi (2018). Expression and characterization of a lysine rich protein in cow milk. Indian Journal of Animal Research. 53(7): 874-879. doi: 10.18805/ijar.B-882.

ABSTRACT

The lysine is considered as the most important essential amino acid, because it is the most limiting in the cereals grains. In this study, a lysine-rich (LR) gene, and the expression vector pcDNA3.1-LR and pBC1-LR were constructed. The LR was expressed in 293T cells driven by the vector pcDNA3.1-LR and checked by RT-PCR and WB. The mammary gland tissue-specific expression vector carrying the LR was injected directly into the lactating mammary glands of cows and the milk samples were checked by a complete amino acid analysis. The results showed that the LR protein was expressed successfully in cells and in cow milk; the expression of LR lasted for 6 d, and the lysine level of the injection group was significantly higher than that of negative controls (p <0.05). This study provide a better understanding of how mammary gland expression systems increase the lysine content of milk that can be applied to transgenic dairy cow.

REFERENCES

  1. Appuhamy, J.A., Knapp, J.R., Becvar, O., Escobar, J., Hanigan, M.D. (2011). Effects of jugular-infused lysine, methionine, and branched-chain amino acids on milk protein synthesis in high-producing dairy cows. Journal of Dairy Science, 94: 1952-1960. 
  2. Brophy, B., Smolenski, G., Wheeler, T., Wells, D., L, H.P., Laible, G. (2003). Cloned transgenic cattle produce milk with higher levels of beta-casein and kappa-casein. Nature Biotechnology, 21: 157-162.
  3. Bicar, E.H., Woodman-Clikeman, W., Sangtong, V., Peterson, J.M., Yang, S.S., Lee, M., Scott, M.P. (2008). Transgenic maize endosperm containing a milk protein has improved amino acid balance. Transgenic Research, 17: 59-71.
  4. Dizigan, M.A., Kelly, R.A., Voyles, D.A., Luethy, M.H., Malvar, T.M., Malloy, K.P. (2007). High lysine maize compositions and event LY038 maize plants. Monsanto Technology LLC, St. Louis, MO(US), assignee. U.S. Patent, 7157281.
  5. DePeters, E.J., Cant, J.P. (1992). Nutritional factors influencing the nitrogen composition of bovine milk: a review. Journal of Dairy Science, 75:2043-2070.
  6. Echelard, Y., Williams, J.L., Destrempes, M.M., Koster, J.A., Overton, S.A., Pollock, D.P., Rapiejko, K.T., Behboodi, E et al ., Meade. production of recombinant albumin by a herd of cloned transgenic cattle. Transgenic Research, 18: 361-376.
  7. Frizzi, A., Huang, S., Gilbertson, L.A., Armstrong, T.A., Luethy, M.H., Malvar, T.M. (2008). Modifying lysine biosynthesis and catabolism in corn with a single bifunctional expression/silencing transgene cassette. Plant Biotechnology Journal, 6: 13-21.
  8. Guenoune, D., Amir, R., Badani, H., Wolf, S., Galili, S. (2003). Coexpression of the soybean vegetative storage protein beta subunit (S-    VSPbeta) either with the bacterial feedback-insensitive dihydrodipicolinate synthase or with S-VSPalpha stabilizes the S-    VSPbeta transgene protein and enhances lysine production in transgenic tobacco plants. Transgenic Research, 12: 123-126.
  9. Jost, B., Vilotte, J.L., Duluc, I., Rodeau, J.L., Freund, J.N. (1999). Production of low-lactose milk by ectopic expression of intestinal lactase in the mouse mammary gland. Nature Biotechnology, 17: 160-164.
  10. Kao, B.T., Lewis, K.A., DePeters, E.J., Van, E.A. (2006). Endogenous production and elevated levels of long-chain n-3 fatty acids in the milk of transgenic mice. Journal of Dairy Science, 89:3195-3201.
  11. Ma, X., Zhang, P., Song, G., Chen, Y., Wang, Z., Yin, Y., Kong, D., Zhang, S., Zhao, Z., Ouyang, H., Tang, B., Li, Z. (2012). The construction and expression of lysine-rich gene in the mammary gland of transgenic mice. DNA and Cell Biology, 31: 1372-1383. 
  12. Persuy, M.A., Legrain, S., Printz, C., Stinnakre, M.G., Lepourry, L., Brignon, G., Mercier, J.C. (1995). High-level, stage- and mammary-    tissue-specific expression of a caprine kappa-casein-encoding minigene driven by a beta-casein promoter in transgenic mice. Gene, 165: 291-296.
  13. Roberts, B., DiTullio, P., Vitale, J., Hehir, K., Gordon, K. (1992). Cloning of the goat beta-casein-encoding gene and expression in transgenic mice. Gene, 121: 255-262.
  14. Soler, E., Le, S.A., Guinut, F., Passet, B., Cohen, R., Merle, C., Charpilienne, A., Fourgeux, C., Sorel, V., et al (2005). Production of two vaccinating recombinant rotavirus proteins in the milk of transgenic rabbits. Transgenic Research, 14: 833-844.
  15. Soler, E., Thepot, D., Rival-Gervier, S., Jolivet, G., Houdebine, L.M. (2006). Preparation of recombinant proteins in milk to improve human and animal health. Reproduction Nutrition Development, 46: 579-588.
  16. Vyas, D., Erdman, R.A. (2009). Meta-analysis of milk protein yield responses to lysine and methionine supplementation. Journal of Dairy Science, 92:5011-5018.
  17. Vilotte, J.L. (2002).Lowering the milk lactose content in vivo: potential interests, strategies and physiological consequences. Reproduction Nutrition Development, 42: 127-132.
  18. Wang, C., Liu, H.Y., Wang, Y.M., Yang, Z.Q., Liu, J.X., Wu, Y.M., Yan, T., Ye, H.W. (2010). Effects of dietary supplementation of methionine and lysine on milk production and nitrogen utilization in dairy cows. Journal of Dairy Science, 93: 3661-3670.
  19. Xu, S., Harrison, J.H., Chalupa, W., Sniffen, C., Julien, W., Sato, H., Fujieda, T., Watanabe, K., Ueda, T., Suzuki, H. (1998). The effect of ruminal bypass lysine and methionine on milk yield and composition of lactating cows. Journal of Dairy Science, 81:1062-1077.
  20. Yang, P., Wang, J., Gong, G., Sun, X., Zhang, R., Du, Z., Liu, Y., Li, R., Ding, F., Tang, B., Dai, Y., Li, N. (2008). Cattle mammary bioreactor generated by a novel procedure of transgenic cloning for large-scale production of functional human lactoferrin. Plos One, 3: e3453.
  21. Yu, Z., Meng, Q., Yu, H., Fan, B., Yu, S., Fei, J., Wang, L., Dai, Y., Li, N. (2006). Expression and bioactivity of recombinant human lysozyme in the milk of transgenic mice. Journal of Dairy Science, 89: 2911-2918. 
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 7 (july 2019) : 874-879,   Doi: 10.18805/ijar.B-882

    Expression and characterization of a lysine rich protein in cow milk

    X
    Xin Ma
    L
    Li Su
    P
    Peng Zhang
    S
    Sheng Zhang
    B
    Bo Tang
    X
    Xueming Zhang
    W
    Weimin Luan
    Z
    Ziyi Li
    1College of Animal Science and Technology, Jilin Agricultural University, Changchun, China.
    Cite article:- Ma Xin, Su Li, Zhang Peng, Zhang Sheng, Tang Bo, Zhang Xueming, Luan Weimin, Li Ziyi (2018). Expression and characterization of a lysine rich protein in cow milk. Indian Journal of Animal Research. 53(7): 874-879. doi: 10.18805/ijar.B-882.

    ABSTRACT

    The lysine is considered as the most important essential amino acid, because it is the most limiting in the cereals grains. In this study, a lysine-rich (LR) gene, and the expression vector pcDNA3.1-LR and pBC1-LR were constructed. The LR was expressed in 293T cells driven by the vector pcDNA3.1-LR and checked by RT-PCR and WB. The mammary gland tissue-specific expression vector carrying the LR was injected directly into the lactating mammary glands of cows and the milk samples were checked by a complete amino acid analysis. The results showed that the LR protein was expressed successfully in cells and in cow milk; the expression of LR lasted for 6 d, and the lysine level of the injection group was significantly higher than that of negative controls (p <0.05). This study provide a better understanding of how mammary gland expression systems increase the lysine content of milk that can be applied to transgenic dairy cow.

    REFERENCES

    1. Appuhamy, J.A., Knapp, J.R., Becvar, O., Escobar, J., Hanigan, M.D. (2011). Effects of jugular-infused lysine, methionine, and branched-chain amino acids on milk protein synthesis in high-producing dairy cows. Journal of Dairy Science, 94: 1952-1960. 
    2. Brophy, B., Smolenski, G., Wheeler, T., Wells, D., L, H.P., Laible, G. (2003). Cloned transgenic cattle produce milk with higher levels of beta-casein and kappa-casein. Nature Biotechnology, 21: 157-162.
    3. Bicar, E.H., Woodman-Clikeman, W., Sangtong, V., Peterson, J.M., Yang, S.S., Lee, M., Scott, M.P. (2008). Transgenic maize endosperm containing a milk protein has improved amino acid balance. Transgenic Research, 17: 59-71.
    4. Dizigan, M.A., Kelly, R.A., Voyles, D.A., Luethy, M.H., Malvar, T.M., Malloy, K.P. (2007). High lysine maize compositions and event LY038 maize plants. Monsanto Technology LLC, St. Louis, MO(US), assignee. U.S. Patent, 7157281.
    5. DePeters, E.J., Cant, J.P. (1992). Nutritional factors influencing the nitrogen composition of bovine milk: a review. Journal of Dairy Science, 75:2043-2070.
    6. Echelard, Y., Williams, J.L., Destrempes, M.M., Koster, J.A., Overton, S.A., Pollock, D.P., Rapiejko, K.T., Behboodi, E et al ., Meade. production of recombinant albumin by a herd of cloned transgenic cattle. Transgenic Research, 18: 361-376.
    7. Frizzi, A., Huang, S., Gilbertson, L.A., Armstrong, T.A., Luethy, M.H., Malvar, T.M. (2008). Modifying lysine biosynthesis and catabolism in corn with a single bifunctional expression/silencing transgene cassette. Plant Biotechnology Journal, 6: 13-21.
    8. Guenoune, D., Amir, R., Badani, H., Wolf, S., Galili, S. (2003). Coexpression of the soybean vegetative storage protein beta subunit (S-    VSPbeta) either with the bacterial feedback-insensitive dihydrodipicolinate synthase or with S-VSPalpha stabilizes the S-    VSPbeta transgene protein and enhances lysine production in transgenic tobacco plants. Transgenic Research, 12: 123-126.
    9. Jost, B., Vilotte, J.L., Duluc, I., Rodeau, J.L., Freund, J.N. (1999). Production of low-lactose milk by ectopic expression of intestinal lactase in the mouse mammary gland. Nature Biotechnology, 17: 160-164.
    10. Kao, B.T., Lewis, K.A., DePeters, E.J., Van, E.A. (2006). Endogenous production and elevated levels of long-chain n-3 fatty acids in the milk of transgenic mice. Journal of Dairy Science, 89:3195-3201.
    11. Ma, X., Zhang, P., Song, G., Chen, Y., Wang, Z., Yin, Y., Kong, D., Zhang, S., Zhao, Z., Ouyang, H., Tang, B., Li, Z. (2012). The construction and expression of lysine-rich gene in the mammary gland of transgenic mice. DNA and Cell Biology, 31: 1372-1383. 
    12. Persuy, M.A., Legrain, S., Printz, C., Stinnakre, M.G., Lepourry, L., Brignon, G., Mercier, J.C. (1995). High-level, stage- and mammary-    tissue-specific expression of a caprine kappa-casein-encoding minigene driven by a beta-casein promoter in transgenic mice. Gene, 165: 291-296.
    13. Roberts, B., DiTullio, P., Vitale, J., Hehir, K., Gordon, K. (1992). Cloning of the goat beta-casein-encoding gene and expression in transgenic mice. Gene, 121: 255-262.
    14. Soler, E., Le, S.A., Guinut, F., Passet, B., Cohen, R., Merle, C., Charpilienne, A., Fourgeux, C., Sorel, V., et al (2005). Production of two vaccinating recombinant rotavirus proteins in the milk of transgenic rabbits. Transgenic Research, 14: 833-844.
    15. Soler, E., Thepot, D., Rival-Gervier, S., Jolivet, G., Houdebine, L.M. (2006). Preparation of recombinant proteins in milk to improve human and animal health. Reproduction Nutrition Development, 46: 579-588.
    16. Vyas, D., Erdman, R.A. (2009). Meta-analysis of milk protein yield responses to lysine and methionine supplementation. Journal of Dairy Science, 92:5011-5018.
    17. Vilotte, J.L. (2002).Lowering the milk lactose content in vivo: potential interests, strategies and physiological consequences. Reproduction Nutrition Development, 42: 127-132.
    18. Wang, C., Liu, H.Y., Wang, Y.M., Yang, Z.Q., Liu, J.X., Wu, Y.M., Yan, T., Ye, H.W. (2010). Effects of dietary supplementation of methionine and lysine on milk production and nitrogen utilization in dairy cows. Journal of Dairy Science, 93: 3661-3670.
    19. Xu, S., Harrison, J.H., Chalupa, W., Sniffen, C., Julien, W., Sato, H., Fujieda, T., Watanabe, K., Ueda, T., Suzuki, H. (1998). The effect of ruminal bypass lysine and methionine on milk yield and composition of lactating cows. Journal of Dairy Science, 81:1062-1077.
    20. Yang, P., Wang, J., Gong, G., Sun, X., Zhang, R., Du, Z., Liu, Y., Li, R., Ding, F., Tang, B., Dai, Y., Li, N. (2008). Cattle mammary bioreactor generated by a novel procedure of transgenic cloning for large-scale production of functional human lactoferrin. Plos One, 3: e3453.
    21. Yu, Z., Meng, Q., Yu, H., Fan, B., Yu, S., Fei, J., Wang, L., Dai, Y., Li, N. (2006). Expression and bioactivity of recombinant human lysozyme in the milk of transgenic mice. Journal of Dairy Science, 89: 2911-2918. 
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